Enhancing tylosin production by combinatorial overexpression of efflux,SAM biosynthesis,and regulatory genes in hyperproducing Streptomyces xinghaiensis strain
作者机构:State Key Laboratory of Microbial MetabolismSchool of Life Sciences and BiotechnologyShanghai Jiao Tong UniversityShanghai200240China Zhejiang Apeloa Biotechnology Co.Ltd.Jinhua322109China Zhejiang Apeloa Jiayuan Pharmaceutical Co.Ltd.Jinhua322118China Haihe Laboratory of Synthetic BiologyTianjin300308China
出 版 物:《Synthetic and Systems Biotechnology》 (合成和系统生物技术(英文))
年 卷 期:2023年第8卷第3期
页 面:486-497页
核心收录:
学科分类:0710[理学-生物学] 071001[理学-植物学] 07[理学]
基 金:the National Key Research and Development Program of China(grant no.2022YFC210540303) the“Major Project”of Haihe Laboratory of Synthetic Biology(22HHSWSS00001)
主 题:Streptomyces xinghaiensis Tylosin Combinational metabolic engineering
摘 要:Tylosin is a 16-membered macrolide antibiotic widely used in veterinary medicine to control infections caused by Gram-positive pathogens and *** improve the fermentation titer of tylosin in the hyperproducing Streptomyces xinghaiensis strain TL01,we sequenced its whole genome and identified the biosynthetic gene cluster *** of the tylosin efflux gene tlrC,the cluster-situated S-adenosyl methionine(SAM)synthetase gene metK_(cs),the SAM biosynthetic genes adoK_(cs)-metFcs,or the pathway-specific activator gene tylR enhanced tylosin production by 18%,12%,11%,and 11%in the respective engineered strains TLPH08-2,TLPH09,TLPH10,and ***-overexpression of metK_(cs)and adoK_(cs)-metFcs as two transcripts increased tylosin production by 22%in the resultant strain TLPH11 compared to that in ***,combinational overexpression of tlrC,metK_(cs),adoK_(cs)-metFcs,and tylR as four transcripts increased tylosin production by 23%(10.93g/L)in the resultant strain TLPH17 compared to that in ***,a negligible additive effect was displayed upon combinational overexpression in TLPH17 as suggested by the limited increment of fermentation titer compared to that in *** analyses indicated that the expression of tlrC and three SAM biosynthetic genes in TLPH17 was considerably lower than that of TLPH08-2 and *** on this observation,the five genes were rearranged into one or two operons to coordinate their overexpression,yielding two engineered strains TLPH23 and TLPH24,and leading to further enhancement of tylosin production over *** particular,the production of TLPH23 reached 11.35 g/*** findings indicated that the combinatorial strategy is a promising approach for enhancing tylosin production in high-yielding industrial strains.